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NC1200: Regulation of Photosynthetic Processes

Annual/Termination Reports (SAES-422): [12/29/2012]

Date of Annual Report: 12/29/2012

Report Information:
  • Annual Meeting Dates: 11/16/12 to 11/18/12
  • Period the Report Covers: 10/2011 to 09/2012

    • Participants:
    Brief Summary of Minutes of Annual Meeting:
    Agenda Of Meeting:

    Friday, Nov. 16, 2012 Evening 19:30 Meet in El Dorado Hotel Lobby (Transport by minivan) 20:00 Dinner at La Famiglia, 22:00 Return to El Dorado Hotel (Transport by minivan)

    Saturday, Nov. 17, 2012 Morning 7:45 Meet in El Dorado Hotel Lobby (Transport by minivan); 8:00 Breakfast in Fleischmann Agriculture Hall (FA), room 207.

    8:30 Announcements and introductions, Jeff Harper 8:45 Welcome by NV AES Interim Dean and Director Ron Pardini. 9:00 Mike Salvicci, USDA ARS Arizona; 9:45 Steve Huber, USDA-ARS, Illinois; 10:30 Break; 10:45 Robert Aiken, Kansas AES; 11:30 Grant Cramer Nevada AES (Guest).

    Afternoon 12:15 Lunch and Business Meeting I, FA 207

    12:30 pm Christoph Benning, MSU AgBioResearch; 14:15 Julie Stone, Nebraska AES; 15:00 Break; 15:15 Don Weeks, Nebraska AES; 16:00 Jeff Harper Nevada AES; 16:45 John Cushman Nevada AES; 17:30 Business Meeting II

    Minutes of Business Meeting:

    1. The renewal proposal was funded effective October 1, 2012. Therefore, this was the first meeting of the NC-1200 group, hosted by Jeff Harper and John Cushman (Nevada AES).

    2. Julie Stone, Bob Spreitzer, and Don Weeks (Nebraska AES) will co-host the Year 2 annual meeting in 2013 with Marty Spalding and Steve Rodermel (Iowa AES) in Omaha, NE (most likely) or at ISU or UNL (depending on financial feasibility).

    3. Christoph Benning has officially taken over as Administrative Advisor to replace Irwin Goldman, and will be supported by MSU AgBioResearch Director Steve Pueppke.

    4. There was lengthy discussion regarding the inclusion of guests, either as potential new participants or graduate students and postdoctoral researchers at the home institution. The general consensus was that participants were welcome, as this serves as an excellent opportunity for the individuals at the hosting institution. The question of confidentiality was raised, as many investigators present new, unpublished work. It was agreed that information presented at the meetings should be considered confidential, and this policy should be made clear to all guests. However, our written annual reports submitted to CRIS are public, which should be kept in mind when preparing reports.

    5. Rob Aiken (Kansas AES) agreed to host the Year 3 meeting in 2014. However, further discussion resulted in the decision to hold the actual meeting in Phoenix, AZ with Mike Salvucci (USDA-ARS, Arizona) aiding in the arrangements. The rationale was to try holding the meeting at a location that has more convenient airline connections to make it easier for members of the group to fly in and out for a short meeting. Regardless of the location, Rob will be responsible for the annual report and the final report.

    6. Possible new participants were also discussed, including Maria Salas-Fernandez (Iowa AES) and Michael Giroux (Montana AES). Both will be invited to present their research at next years annual meeting.

    7. Long-standing collaborations among the members were noted and are apparent from joint publications and funding. Many new collaboration opportunities were discussed. As the makeup of the group evolves, current members were strongly encouraged to cement these relationships within the group.

    Accomplishments:
    Accomplishments Summary of Accomplishments Presented by Meeting Attendees

    Objective 1. Plastid Function and Intracellular Communication

    Steve Rodermel (Iowa AES) reported in writing about his labs recent characterization variegation mutants in Arabidopsis as a model system for understanding photosynthetic electron flow. Suppressor lines to the immuntans mutant, which is deficient in PTOX (plastid terminal oxidase), a versatile plastoquinol (terminal) oxidase in plastid membranes, are in various stages of characterization. One activation-tagged suppressor that rescues immutans encodes a mitochondrial AOX2 that is localized to the plastid, specifically in the thylakoid and is able to specifically rescue the phytoene accumulation defect of immutans and is thought to function by supplementing PTOX activity during the early stages of chloroplast biogenesis. The work highlights the important role of PTOX during normal chloroplast biogenensis, chlororespiration, and light-to-dark transitions to poise the redox state of the electron transport chain.

    Mike Salvucci (Arizona ARS) described in his oral presentation a series of studies on improving RUBISCO Activase to achieve higher thermotolerance of photosynthesis. He reported that RUBISCO activase has been crystallized and its structure solved except for missing N and C-terminal regions of the protein. He reported on a series of experiments that determined the mechanisms of redox regulation of RUBISCO activase isoforms. The alpha form is redox regulated, whereas the beta form is not. In species such as Arabidopsis and Camelina, which express alpha and beta forms of the enzyme, physiological ratios of ADP/ATP cause significant inhibition of Rubisco activase activation. Similarly, tobacco, which expresses only the beta form, is also inhibited. However, the tobacco beta form was marked more sensitive to ADP inhibition than the Arabidopsis beta isoform confirming that these two isoforms are differentially regulated by ADP/ATP ratios or when associated with an alpha form by ADP/ATP ratio in combination with changes in redox state. He also reported that Arabidopsis plants expressing a Rubisco activase enzyme that is not inhibited by ADP and thus not deactivated by low irradiance, showed no lag in CO2 fixation during the transition from low to high irradiance, but that elimination of this lag did not lead to increased productivity in fluctuating light environments. Collaborations are ongoing between the Salvucci group and Robert Spreitzer (AES, Nebraska) and Rebekka Wachter (Arizona State University) to investigate the interaction between Rubisco activase and Rubisco using Chlamydomonas Rubisco mutants.

    Julie Stone (Nebraska AES) presented an oral report on her recent work in elucidating the functional roles of three Arabidopsis homologues to the Human DJ-1 protein, which functions as an obligate homodimer to protect against reactive oxygen species-mediated programmed cell death. This functionality is dependent upon a highly conserved redox-sensitive cysteine residue. The redox-sensitive cysteine is conserved in two of the Arabidopsis proteins (AtDJ1A and AtDJ1B), but it is substituted in the third gene (AtDJ1C), which is also predicted to have an N-terminal chloroplast transit peptide. She also reported that Arabidopsis contains three additional more distantly related genes belonging to the DJ-1 superfamily. Her group has now identified T-DNA insertion knockout mutant for the plastid localized AtDJ1C gene product in A. thaliana. Mutant alleles of this gene confer an albino, seedling-lethal phenotype indicating that at least one DJ-1 gene is essential for viability.

    Objective 2. Photosynthetic Capture and Photorespiratory Release of CO2

    James Moroney (Louisiana AES) reported in writing about his labs recent characterization of the carbonic anhydrase gene family in Chlamydomonas. Of the nine gene family members, two of the protein products were found to localize in the chloroplast thylakoid lumen (CAH3) and the stroma (CA6). While both genes were expressed constitutively and their expression enhanced by limiting CO2 concentrations, only CAH3 appears to be an essential component of the carbon concentrating mechanism (CCM). In addition, insertional mutagenesis screens were used to identify a novel protein, CIA6, which has weak homology with RUBISCO-interacting proteins and functions in pyrenoid formation and growth under conditions of limiting CO2. Additional insertional mutagenesis screens have resulted in the isolation of additional mutants that grow slowly on low CO2, but normally on elevated CO2. Lastly, the Moroney lab screened Arabidopsis carbonic anhydrase knockout mutants. One knockout line, with a lesion in beta CA5, shows poor growth under all conditions tested, whereas another lines defective in CA4 shows reduced growth under CO2-limiting conditions. These results are critical to understanding how algae accomplish this uptake of CO2 and how they enhance photosynthesis when the CO2 concentration is low

    Robert Spreitzer (Nebraska AES) reported in writing on various studies elucidating the structural basis of Rubisco catalytic efficiency and CO2/O2 specificity using the Chlamydomonas Rubisco as a model as the structure of this enzyme has been solved to the highest resolution (1.4 Å) of all Rubisco enzymes, and that structures of several mutant enzymes have also been solved. Recent studies have indicated that engineered changes in either the large or small subunit far from the active site can influence carboxylation catalytic efficiency and CO2/O2 specificity and that such distant regions will be targeted for either the design of an improved Rubisco or for genetic selection following mutagenesis. This observation also suggests that combinatorial approaches might be necessary for improving the enzyme. Lastly, two alphas helixes of the Rubisco small subunit are now known to be sufficient for species-specific pyrenoid targeting of Rubisco. Future work might define the specific structural interactions required for establishing the pyrenoid and CO2 concentrating mechanism. Collaborations are ongoing between the Spreitzer group and Mike Salvucci (Arizona ARS) to define the structural interactions between Rubisco and Rubisco activase using Chlamydomonas Rubisco mutants. Spreitzer has generated hybrid Rubiscos (Chlamydomonas RBCL, higher plant RBCS) that have been analyzed and he is now producing alterations at the C-terminus of the RBCL for the analysis of the interaction with Rubisco activase

    Don Weeks (Nebraska AES) presented an oral report on his collaborative work with Martin Spalding (Iowa ARS) and Bing Yang, also at Iowa State University, on the development of TAL Effector Nuclease (TALEN) technology for targeted gene knockout in eukaryotic organisms. Success by this set of collaborators with use of TALEN technology for gene knockout and gene replacement by homologous recombination in yeast cells was described as an example of the power and ease with which the TALEN system can be employed. As a demonstration of the power of the technology for practical application in agriculture, Weeks described experiments conducted primarily in the laboratory of Bing Yang in which a disease susceptibility gene, Os11N3, in rice was targeted by two independent sets of TALENs for knockout. Both sets of TALENs proved successful in causing inactivation of the disease susceptibility gene and the production of rice plants resistant to bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo). Genetic crosses between disease resistant plants allowed the selection of disease resistant plants that lacked the TALE gene set, thus producing rice plants that no longer contained foreign genetic elements. The potential application of this technology to allow targeted gene knockout in other higher plants and in algae was discussed as a means of improving photosynthetic carbon capture and photosynthetic efficiencies.

    Objective 3: Mechanisms Regulating Photosynthate Partitioning

    Christoph Benning (MSU AgBioResearch) provided an oral report updating his recent efforts to increase the production of triacylglycerols (TAGs) in plant biofuel feedstocks given that the need for biodiesel for trucking and aviation and the inability of current feedstocks, such as soy, to meet current and future demands without competing for dietary oils. He summarized results of increasing oil biosynthesis in vegetative tissues of Arabidopsis by over expression of the WRI transcription factor from Arabidopsis as well as with repression of ADP-glucose pyrophosphorylase by RNAi resulted in up to 5% TAG accumulation by dry weight in Brassica napus leaf tissues. Feeding plants with 3% sucrose can result in 10% TAG accumulation, but the plants showed aberrant growth. He also reported on the identification and characterization of Diacylglycerol Acyltransferase Type Two (DGTT) enzymes from Chlamydomonas reinhardtii. Overexpression of DGTT2 in Arabidopsis increase TAG content of leaves along with the production of very long chain fatty acids (C27-C33 alkanes). In addition, acyl carbon partitioning in plant vegetative tissues was altered with increased sphingolipid accumulation and reduced surface wax and cutin accumulation. When caterpillar herbivores (Spodoptera exigua) were reared on the transgenic plants, the insects gained more weight relative to control plants, demonstrating an increased energy content and energy density of the transgenic lines. Lastly, results were presented using wild type and mutants of the model alga Chlamydomonas reinhardtii to investigate the mechanistic basis of the accumulation of TAGs in response to nitrogen deprivation. On mutant tentatively designated Plastid Galactoglycerolipid Degradation 1 (pgd1), defective in the expression of a galactoglycerolipid lipase-encoding gene, exhibited reduced TAG content, altered TAG composition, and reduced galactoglycerolipid turnover. In vivo pulse-chase labeling studies revealed that galactoglycerolipid pools serve as major source of fatty acids esterified in triacylglycerols following N deprivation and that the fatty acid flux from plastid lipids to triacylglycerol was decreased in the pgd1 mutant. In addition, the pgd1 mutant exhibited a loss of viability following N deprivation, which could be avoided by blocking photosynthetic electron transport, indicating that triacylglycerol synthesis following N deprivation functions to relieve a detrimental over reduction of the photosynthetic electron transport chain.

    Jyan-Chyun Jang (Ohio AES) reported in writing about his laboratories research on the molecular mechanisms underlying sugar and hormone sensing and signal transduction in plants. His group investigated the functional roles of the CCCH Tandem Zinc Finger (TZF) family of transcription factors such as AtTZF4, 5, and 6, which are positively regulated by ABA and negatively regulated by GA and themselves can act as positive and negative regulators of ABA and GA accumulation and response, respectively. Compared to the wildtype, knockout lines germinate faster, whereas over-expression lines are delayed in seed germination. Like AtTZF1, over-expression of AtTZF4, 5, and 6 causes compact rosette and enhanced stress tolerance phenotypes. Consistent with the hypotheses that TZF proteins play pivotal roles in the assembly of messenger ribonucleoprotein (mRNP) granules, such as Processing Bodies (PBs) and Stress Granules (SGs) that are hubs for the regulation of gene expression at the post-transcriptional level, AtTZF4, 5, and 6 were found to co-localize with PB and SG markers and are thought to play roles in nucleating these structures by binding to AU-rich elements at the 3UTR, and recruit decapping, deadenylation, and exonucleolytic enzymes to PBs for mRNA turnover as is observed in mammalian systems. Specifically, AtTZF4, 5, and 6 might function in seed-specific ABA/GA response by PB- and SG- mediated post-transcriptional regulation of gene expression. Lastly, the group has conducted a forward genetic screen to identify mutants with revertant phenotypes in an AtTZF1 over-expression background. To date, three classes of mutants have been identified and work is in progress to genetically and functionally characterize these mutants.

    Glenda Gillaspy (Virginia AES) reported in writing that her group investigated the mechanisms that regulate sugar sensing and photosynthate partitioning into biosynthetic pathways. The sucrose non-fermenting related kinases (SnRKs) are major energy sensors in eukaryotes that impact global transcription programs, stress signaling and lifespan. Work using transient expression in Nicotiana benthamiana to measure protein binding within the SnRK complex was reported. A novel protein (P80) was identified that is associated with the ubiquitin-ligase pathway and might act to protect SnRK1.1 from proteosomal destruction. Genes involved in the control of inositol oxidation were also investigated. Two genes, myo-inositol oxygenase (MIOX) 2 and 4, were found to be repressed in shoots, but not roots, by exogenous glucose application. Loss of function mutant analysis revealed that MIOX catabolism in shoots might influence root growth responses under low energy/nutrient conditions.

    Steve Huber (Ilinois ARS) provided an oral report on his work highlight novel aspects of calcium-dependent protein kinases (CPKs), particularly the ability of this class of protein kinase (e.g., AtCPK4 and AtCPK34) to autophosphorylate Tyr residues, confirming that they possess dual specificity. He indicated that dual specificity is common in other classes of protein kinases, where about 40% of receptor kinases will phosphorylation both Ser/Thr and Tyr residues, however, this activity is rare in CPKs. CPK-beta autophosphorylates at a single Tyr site at the N-terminal side of the kinase domain. Removal of this Tyr residue results in greater autophosphorylation and enzyme activity and that calcium appears to inhibit the accessibility of this site. Future research into the specific roles CPKs function via phosphotyrosine signaling events is being investigated in collaboration with the Harper lab. He also reported the novel observation that calmodulin (CaM) can bind in vitro to a predicted high affinity CaM binding site within the variable N-terminal domain of CPK-beta from soybean. He noted that while predicted CaM binding sites are common among CPKs, this is the first reported documentation of such binding. Some CaM isoforms binding can block Tyr autophosphorylation, however, such binding does not appear to inhibit kinase activity in general. Lastly, he described a new E. coli-based transphosphorylation assay to monitor protein kinase specificity. Bacterial protein substrate phosphorylation events appear to correlate with the intrinsic substrate specificity of the kinase assayed, at least for Thr and Tyr sites.

    Karen Koch (Florida AES) reported in writing on her recent development of public resources of UniformMu mutant collection for approximately 40% of the maize genome. A particular emphasis of the Koch lab is the isolation and genotypic and phenotypic characterization of mutants that affect photosynthate partitioning and sugar signaling. Her lab is also charactering the role of strigolactones (SLs) in controlling lateral branching by regulation of the Tb1 transcription factor. Knockout of Carotenoid Cleavage Dioxygenase 8 (CCD8), a gene essential for SL biosynthesis, results in increase branching, reduced stem diameter, reduced elongation of internodes, and developmental delay of the nodal system of adventitious roots. Analysis of double mutants of ccd8 and tb1 indicates that in maize, the SL signaling network appears to function independently of the Tb1 network and accounts for diverse aspects of plant architecture. Lastly, her lab has investigated the functional roles of the cellulose-synthase-like-d group (CslD) of genes within the cellulose-synthase gene superfamily. Knockout of the cellulose-synthase-like-1 (csld1) gene resulted in plants that accumulated 50% less biomass arising mainly from a narrow-organ phenotype that resulted from ineffective cross-wall formation early in organogenesis. These results showed that early-development partitioning of photosynthate to cell wall material results in long-term implications for biomass accumulation patterning later in development.

    Tom Sharkey (MSU AgBioResearch) reported in writing that progress was made on understanding the feedback from triose phosphate use on photosynthetic electron transport and Calvin-Benson cycle activity. A method for measuring partitioning between starch and sucrose was established and it was shown that phosphoglucomutase (pgm) mutants had little to no carbon flux to starch as predicted, but triose phosphate translocator (tpt) mutant plants had relatively more sucrose than predicted. New hypotheses about carbon export from chloroplasts during the day are under development to explain this phenomenon. This work will provide needed information to lift the limits on photosynthesis imposed by feedback effects and will show how sucrose/starch partitioning in photosynthesis affects plant development through sugar signaling.

    Objective 4: Developmental and Environmental Limitations to Photosynthesis

    Rob Aiken (Kansas AES) summarized in his oral presentation a 2008 World Development Report that anticipated large increases in drought-associated evapotranspiration rates based on increasing surface temperatures. Above ground biomass is more or less linear with crop water use in the 200-500 mm precipitation range, but falls off below 200 mm. Dr. Aiken then results of conducted by Vara Prassad on screening of 52 accessions of wild wheat germ plasm for heat tolerance and found that Aegilops speltoides and A. geniculate accessions were most tolerant. When Aegilops speltoides was used as the female parent to create hybrids or synthetic hexaploids, these lines showed the least chlorophyll loss under heat (and drought) stress conditions. Further results of different sorghum genotypes identified lines with differences in their sensitivity to elevated vapor pressure deficit and revealed different strategies for soil water conservation in water-deficit environments existed. One group exhibited reduced transpiration at a higher threshold of the fraction of transpirable soil water (FTSW), whereas another group exhibited low transpiration rates at all times, resulting in conservation of soil water content at all times. Lastly, leaf structure was studied in five grass species, including C3 and C4 functional types, and indicated that both stomatal conductance and photosynthesis increased along all grass blades despite constant light availability.

    Fred Below and Laura Gentry (Illinois AES) reported in writing on their development of a more efficient method for breeders to use to determine a maize plants potential yield response to nitrogen fertilizer. In their studies with temperate x tropical maize (TTM), TTM partitioned 50% more biomass to the stalk, produced 50% more sugar, and had less than half the grain of the commercial hybrids, indicating that grain production and sugar accumulation are inversely related. Calculated estimates for ethanol production, considering the potential from sugar, stover and grain, indicate that TTM can yield nearly the amount of ethanol per hectare as modern grain hybrids, but with a decreased requirement for supplemental fertilizer N. In their investigation of corn stover residue and C/N balance, continuous corn treatments produced, on average, 13 bu a-1 less than corn-soybean rotations. Continuous corn rotations sequestered an average of 62% more C in intermediate- and long-term soil carbon pools than corn-soybean rotations. Stover removal had a positive effect on corn yield in continuous corn systems, especially under increased plant density with advanced N and non-N fertility levels. Strip tillage performed as well as conventional tillage in corn-soybean rotations, but resulted in yield loss in continuous corn systems. Increasing plant density by 40% increased the effects of drought stress, especially in continuous corn systems. Advanced nutrition (N, P, S, and Zn applications) increased corn yields significantly.

    Jiaxu Li (Mississippi State AgBioResearch) described the analysis of phosphoproteins regulated by abscisic acid-activated protein kinases in Arabidopsis using T-DNA insertion knockout mutants and proteomic tools in his written report. Several phosphoproteins were identified by tandem mass spectrometry. These phosphoproteins could be the substrates of abscisic acid-activated protein kinases in Arabidopsis.

    John Cushman (Nevada AES) described in his oral presentation two different high throughput metabolomic studies using a commercially available platform (Metabolon, Inc.). One study compared two different, but closely related species of Selaginella wherein one species was desiccation tolerant and the other desiccation sensitive. In the sister group comparison, a total of 301 metabolites were identified including 170 known and 131 unnamed compounds. S. lepidophylla retained significantly higher abundances of sucrose, mono- and polysaccharides, and sugar alcohols than did S. moellendorffii. Aromatic amino acids, the well-known osmoprotectant betaine, and flavonoids were also more abundant in S. lepidophylla. In addition, the desiccation tolerant S. lepidophylla contained 14 unnamed compounds of 5-fold or greater abundance than in S. moellendorffii, suggesting that these compounds might play critical roles in desiccation tolerance. He also described a second study in which a total of 251 metabolites were characterized in S. lepidophylla subjected to a five-stage dehydration-rehydration cycle. Only 42 (16.7%) and 74 (29.5%) of compounds showed significantly increased or decreased abundance, respectively, indicating that most compounds were produced constitutively including highly abundant trehalose, sucrose, and glucose. He also reported that many different sugar alcohols and sugar acids were more abundant in the hydrated state that he speculated to have roles in slowing the rate of water loss during drying and slowing the rate of water absorption during rehydration, stabilizing proteins, and scavenging reactive oxygen species (ROS). A set of g-glutamyl amino acids, citrulline, and nucleotide catabolism products (e.g., allantoin) were more abundant in the dry states, suggesting that these compounds play important roles in nitrogen remobilization during rehydration or in ROS scavenging. Lastly, a series of potent antioxidant or UV-protective or antioxidant compounds, such as 3-(3-hydroxyphenyl)propionate, apigenin and naringenin, were described as being more abundant in the dry states. These results indicated that desiccation tolerance requires the combination of both constitutively expressed and inducible metabolites.

    Jeff Harper (Nevada AES) reported by way of his oral presentation on novel aspects of Cyclic Nucleotide Gated Channels (CNGCs) function. These ion channels have been implicated in calcium signaling and are regulated by both cyclic nucleotides and calmodulin. He reported that at least six CNGCs are expressed in pollen of Arabidopsis and that several are essential for pollen viability including CNGC18 and 7/8. In contrast, some are not essential, such as CNGC16, under normal growth conditions. However, under condition where plants are stressed under a cold night and hot day temperature regime, cngc16 mutants show near complete male sterility and an attenuated transcriptional stress response suggesting that stress-triggered cyclic nucleotide and/or calcium signaling via CNGCs might participate in stress adaptive responses. He also reported recent progress on investigations into the function of a family of 12 lipid flippases (ALAs) belonging to a unique subfamily of P-type ATPases. A knockout mutant of one family member, ala3, exhibits reduced germination and growth, especially under middles temperature stress conditions, suggesting that the ALA3 gene product participates in the trans-Golgi network in membrane signalling or structure. A second double knockout mutant, ala6/7, showed reduced pollen tube growth and resultant seed set, suggesting that these genes function in the plasma membrane to regulate the abundance of specific lipids in the membrane under the control of calcium-dependent protein kinases.

    Mike Salvucci (Arizona ARS) also described in his oral presentation a series of studies on improving Rubisco activase to achieve higher thermotolerance of photosynthesis. He reported that by creating thermal denaturation profiles generated by circular dichroism, creosote and tobacco beta-Rubisco activases were among the most stable proteins examined with mid-point temperatures of 45-47 ºC for protein denaturation. He also determined that ADP-bound Rubisco activase was more stable than ATP-bound enzyme and that a two-subunit module provides the basic building block for larger multi-subunit assemblies that typically are smaller than a hexamer. Collaborations are ongoing between the Salvucci group and John Cushman (Nevada AES) and to investigate the thermostability of the Opuntia Rubisco activase, a CAM plant with exceptional thermotolerance, and to introduce heat stable activase into crop plants.

    CURRENT FUNDING Fred Below

    Dudley Smith Initiative. Developing tropical maize as a new biofuel and forage crop for Illinois. $600,000. (05/01/10-04/30/16).

    Christoph Benning US Department of Energy. Great Lakes Bioenergy Research Center. Cooperative Agreement DE-FC02-07ER64494. Multi-investigator project, T. Donahue PI. Funds to my lab ~$250,000 per year based on annual renewal. Regulation of oil biosynthesis in vegetative tissues and the engineering of biofuel crop plants producing oil in roots and shoot tissues. (12/01/12-11/31/17).

    US AFOSR. Regulation of oil biosynthesis in algae. $561,382 (09/01/11-08/31/14).

    NSF MCB 0741395. Mechanisms of lipid trafficking between the endoplasmic reticulum and the chloroplast. $546,818 (05/01/08-04/30/11).

    DOE, DE-FG02-98ER20305. Regulation of Thylakoid Lipid Biosynthesis in Plants. $560,000 (08/01/10-7/31/13).

    DOE, GLBRC. Multi-investigator project, T. Donahue PI. Funds to CB~$200,000 per year based on annual renewal. Regulation of oil biosynthesis in vegetative tissues and the engineering of biofuel crop plants producing oil in roots and shoot tissues. (09/01/07-08/31/12).

    John Cushman NSF IOS-084373. Regulatory and signaling mechanisms of crassulacean acid metabolism: A photosynthetic adaptation of environmental stress. $988,389 (8/15/09-8/14/12).

    NSF DBI 0741876. Research Coordination Network (RCN) Program. RCN: An international research and education collaboration for grape functional genomics. $509,420 (9/15/08-9/14/13).

    Hatch NEV-00372. Improved abiotic stress tolerance of Camelina: A Novel Biofuel Crop for Nevada. PI, Co-PI J. Harper. $529,900 (7/1/10-6/30/15).

    DOE, SBIR Phase II DE-SC0001306. High efficiency microalgae biofuel harvest and extraction using ionic liquids. $60,766 (10/1/10-9/30/12).

    Hatch NEV-00377. Development of Opuntia (Prickly Pear Cactus) as a low water input oleogenic biofuel and biomass feedstock. PI, Co-PI D. Shintani. $157,348 (7/1/12-6/31/17).

    DOE Genomic Sciences Program. Engineering CAM photosynthetic machinery into bioenergy crops for biofuels production in marginal environments. PI, Co-PIs: Karen Schlauch, James Hartwell, and ORNL researchers (A. Borland, J-G Chen, M. Martin, T. Tschaplinski, G. Tuskan, D. Weston, and X. Yang). Total Award $14,356,653, UNR ~$7,632,695 (09/01/12-09/30/18).

    Gerry Edwards

    IRRI (Bill and Melinda Gates Foundation). Creating the Second Green Revolution by supercharging photosynthesis: C4 Rice (PI, part of a consortium under the leadership of IRRI, The Philippines; co-PI A. Cousins). $334,959 (10/01/08-04/31/12).

    CRDF. Identifying mechanisms for increasing carbon acquisition and water use efficiency of plants with climate change. (PI, co-PI A Cousins and Russian investigators) $54,886 (9/1/10-9/1/12).

    USDA. Improving Plant Productivity by Altering Nitrogen Transport Processes. (Co-PI, PI M. Tegeder) $350,000 (12/15/09-12/14/12).

    USDA. Biosecurity Special Research Grant Aegilops cylindrical. (Co-PI, M. Kahn PI). $28,500 funding available to Edwards (Exp. 7/31/12).

    Glenda Gillaspy

    NSF MCB1051646 Collaborative Research: Diphospho- and Triphospho-Inositol Phosphates in Plants $673,426 (1/1/11- 12/31/14)

    NSF Collaborative Research: Diphospho- and Triphospho-Inositol Phosphates in Plants (PI: Gillaspy) Total Award Amount: $673,426. (1/1/11- 12/31/14).

    Steve Huber

    United Soybean Board, BBI Oil Project #7222. $40,000 annual (3/1/10-2/29/13)

    NSF, MCB-1021363. Arabidopsis 2010: Protein interacting networks and site-specific phosphorylation in leucine-rich repeat receptor-like kinase function. Multi-investigator project, S.C. Clouse, PI. Funds to SH lab ~$200,000 per year based on annual renewal (9/1/2010-8/30/2014).

    NSF, IOS-1022177. Regulation of receptor kinase signaling by tyrosine phosphorylation and calmodulin binding. $499,166 (9/01/2010-8/30/2013).

    Jeff Harper

    NSF, MCB-0920624. Calcium Dependent Protein Kinases in Pollen Tube Tip Growth. $390.000 (09/01/09-08/31/12).

    DOE, DE-FG03-94ER20152. P-type ATPases in Plants Role of Lipid Flippases in Membrane Biogenesis $390,000 (09/01/09-08/31/12).

    JC Jang

    Ohio Plant Biotechnology Consortium (2011-005). The novel roles of tandem zinc finger proteins in multi-stress tolerance in plants. $60,000 (07/01/2011-06/30/2013).

    Karen Koch

    USDA-NRI-Plant Biochemistry 07-03580. Roles and regulation of sorbitol metabolism in maize. $394,306 (PI) (09/01/2007-08/31/2012 w/2 NCE).

    NSF-PGRP 08-21952. TRMS: Functional genomics of transfer cells. $1,986,692 (Co-PI with 2 others) (09/01/2008-08/31/2012 w/ NCE).

    NSF-PGRP-IOS 1116561. UniformMu: A transposon resource for mutagenesis in maize. $1,986,692 (2 PIs) (09/01/2011-08/31/2013).

    NSF-PGRP-IOS 1025976. GEPR/LIT Genetic and genomic approaches to understanding long-distance transport and carbon partitioning. $6,100,000 (Co-PI with five others). (12/01/2010 - 11/31/2015).

    USDA-NIFA 2011-67003030215. Adapting kernel metabolism to enhance cereal yield under adverse conditions. $5,000,000 (Co-PI with nine others). (08/01/2011- 07/31/2016).

    David Kramer

    NIH, (2 RO1 GM061904) Rieske Headgroup Cytochrome Complexes - Metal Ion Probes Membrane Protein Structure and Function (Co-PI with Dr. Michael Bowman) $2,057,830 (1/1/09-12/31/12).

    DOE (DE-FG02-04ER15559) The Energy Budget of Steady-State Photosynthesis $540,000 (7/1/08-6/30/12).

    USDA, Plant Biology (C): Biochemistry (2008-35318-04665) Co-regulation of the light and dark reactions of photosynthesis. $400,000 (1/01/09-12/31/12).

    DOE (DE-EE0003046) The National Alliance for Advanced Biofuels and Bioproducts an Algal Biofuels Consortium (Lead Consortium Lead: The Donald Danforth Plant Science Center, Executive Director: Jose Olivares, Los Alamos National Lab and Donald Danforth Plant Science Center). $54,000,000 ($660,000 to MSU) (1/01/10-12/31/13).

    NSF. (RC100150) Plug and Play Photosynthesis for RuBisCO Independent Fuels $3.6M for 7 co-P.I.s, $300,599 to MSU (06/01/2011-05/31/2014).

    DOE. Center for Advanced Camelina Oil (CECO) 9.3M ($860,000 to MSU, in negotiation), (1/1/12-12/31/15).

    Jiaxu Li

    USDA-(NIFA) 2009-04041 Role of histone H3 lysine 36 methylation in regulating developmentally important genes in rice. $134,815 (01/01/10 - 12/31/11).

    Wayne Loescher USDA FAS 58-3148-8-159 Enhancing salinity tolerance in canola $60,000 (9/1/09-8/31/12).

    USDA-CSREES/NIFA 2010-85117-20570 Building Expertise in Plant Breeding that Focuses on Drought Tolerance $500,000 (2/1/10 -1/31/14).

    Steve Rodermel

    DOE, DE-FG02-10ER20147. The immutans variegation mutant of Arabidopsis. $510,000 (10/01/10-7/31/13).

    NSF (UCSD PO 10301097-001). Regulation of chloroplast gene expression. $195,000. (07/01/09-06/30/12) UISFL. Global pathways for educating students in biodiversity. $179,000 (10/01/09- 9/30/12).

    Mike Salvucci

    DOE, DE-AI02-97ER20268. Optimizing Rubisco Regulation for Increased Photosynthetic Performance under Climate Change. $381,000 (05/01/2010-04/30/2012).

    Robert Spreitzer

    DOE, DE-FG02-00ER15044. Role of the Rubisco Small Subunit. $495,000 (05/01/10-04/30/13).

    NSF, EPSCoR. Nebraska Center for Algal Biology and Biotechnology. Multi-investigator project. Funds to my lab ~$306,322 (10/01/10-09/30/15).

    Thomas D. Sharkey

    NSF, IOS-0950574.Function and regulation of isoprene synthesis in leaves. $590,000 (02/15/11-01/31/14).

    ZuvaChem Inc. Novel isoprene synthase genes for isoprene production. $300,000. (07/01/11-06/30/14).

    DOE DE-SCOOO8509. Improved Efficiency of Energy Capture and Conversion By Regulating the Interaction Between ATP Synthesis and End Product Synthesis.

    Julie Stone

    NSF-DBI-0851747. NSF REU SITE: Training in Redox Biology. PI: Don Becker; co-PI: Julie Stone. $252,250 (4/09-3/12).

    NSF, DBI-1156692. NSF REU SITE: Training in Redox Biology. PI: Don Becker; co-PI: Julie Stone. $278,500 (4/12-3/15).

    Univ. Nebraska IANR. Novel Subunits of Serine Palmitoyltransferase, New Players in Sphingolipid Synthesis PI: Ed Cahoon, co-PI: Julie Stone. $60,000 (05/01/2010-04/40/2012).

    Univ. Nebraska IANR. Plant Stress Biology Initiative Co-PIs: Julie Stone, Gary Brewer, $15,000 (05/2010-04/2012).

    Don Weeks

    National Science Foundation MCB-0952533 Bicarbonate Transport in Chlamydomonas $540,000 (3/15/2010  02/28/2013).

    National Science Foundation EPSCoR-1004094 Building Infrastructure in Nanohybrid Materials and Algal Biology Research $412,000 (10/01/2010  09/30/2015).

    Department of Energy DE-EE0001052 Renewable Biofuels from Algae $200,000 (02/01/2010  01/31/2012).

    Department of Energy  CAB-COMM DE-EE0003373 Consortium for Commercialization of Algae Biofuels and Biotechnology $300,000 (09/01/2010  08/31/2013).

    Impact Statements:
    1. Discovery of the target proteins of abscisic acid-activated protein kinases would reveal new components involved in abscisic acid signaling and may provide novel targets for genetic engineering of drought tolerance in crops.
    2. Development of a nitrogen response index to evaluate maize nitrogen use efficiency and increased population using only three nitrogen fertilizer rates.
    3. Grain production and sugar accumulation in maize are inversely related. Calculated estimates for ethanol production, considering the potential from sugar, stover and grain, indicate that temperate x tropical maize hybrids can yield nearly the amount of ethanol per hectare as modern grain hybrids, but with a decreased requirement for supplemental fertilizer N.
    4. Continuous corn rotations sequestered an average of 62% more C in intermediate- and long-term soil carbon pools than corn-soybean rotations. Stover removal had a positive effect on corn yield in continuous corn systems, especially under increased plant density with advanced N and non-N fertility levels.
    5. Discovery of new protein components that may regulate stability of the SnRK1 energy sensor. These proteins are relevant targets for genetic engineering to increase plant fitness and/or biomass.
    6. Recent genetic analyses has indicated that plant TZFs are important for hormone-mediated growth and stress responses, specifically in the formation of Processing Bodies (PBs) and Stress Granules (SGs) that have now been confirmed to exist in plants and appear to act as hubs for the regulation of gene expression via post-transcriptional processing events associated with the sugar and hormone signal transduction.
    7. Screening of germplasm resources of wild wheat relatives and discovery of high temperature tolerance phenotypes, particularly at the reproductive stage, and introgression of this trait into domestic wheat varieties will lead to varieties with improved heat and drought stress tolerance.
    8. Study of leaf structure and function will reveal new relationships between stomatal conductance and density, and photosynthesis and water use efficiency.
    9. Identification of low or differential transpiration rates as it relates to soil moisture conservation in water-deficit environments could lead to the identification of water conservation traits in sorghum grown in dry land environments.
    10. Improved mechanistic understanding of conversion and partitioning of photosynthate into lipids essential for the engineering of novel biofuel crops such as algae or perennial grasses.
    11. Development of plant feedstocks for biodiesel production by increasing TAG content in vegetative structures and increase in the energy density of these structures leading to feedstocks suitable for both lignocellulosic- and TAG-derived biofuels.
    12. Improved accumulation of oil in vegetative tissues could enhance the nutritional and/or caloric content of feed for animals.
    13. Discovery of novel metabolites in resurrection species such as Selaginella resulted in the identification of osmoprotectants, reactive oxygen scavenging metabolites, and UV-protective metabolites that could be used to engineer improved crop tolerance to heat or drought stress.
    14. A detailed evaluation of desiccation tolerant grasses belonging to the Poaceae, such as Sporobolus stapfianus, indicated that Sporobolus species possess forage quality traits (e.g., crude protein, fiber content) comparable to those of timothy, a forage grass grown widely in the Great Basin in the western United States, and that such grasses might serve as useful forage crops in semi-arid and arid regions of the western U.S. where irrigation resources are limited.
    15. SnRK1 energy sensor and myo-inositol levels in the pathways plants use to sense and respond to energy. Manipulation of SnRK1 and myo-inositol levels might be valid targets for genetic engineering to increase plant fitness and/or biomass.
    16. Specific members of the cyclic nucleotide Ca2+-permeable ion channel gene family expressed in pollen grains have been shown to have critical functions in pollen viability, especially under extreme temperature conditions, indicating these ion channel link stress-triggered cyclin nucleotide and calcium signaling events with stress adaptive response pathways.
    17. Specific members of the lipid flippases (ALA) belonging to a unique family of P-type ATPases that function to flip lipids from one membrane leaflet to the other are likely to enable signaling or structural changes in membrane lipids by modulation of the abundance of specific lipid head groups under the control of calcium-dependent protein kinases.
    18. A fundamental understanding of receptor kinase-mediated phosphorylation and the interaction of calmodulin to differentially affect kinase activities has important implications for the regulation of pathogen-associated molecular pattern (PAMP)-triggered immunity in PAMP-triggered immunity in plants.
    19. Novel autophosphoryation on Tyrosine residues was observed for soybean CPK-beta and several Arabidopsis CPKs (AtCPK4 and 34) with an apparent attenuation of kinase activity raising the possibility that phosphotyrosine signaling might play a role in calcium/CPK-mediate signaling processes.
    20. Public mutant collections have been developed covering approximately 40% of the maize genome and has been used to identify mutations that affect photosynthate partitioning and sugar signaling.
    21. Characterization of a knock-out mutation in Carotenoid Cleavage Dioxygenase 8 (CCD8), gene essential for the biosynthesis of strigolactones, which are involved in the control of later branching, revealed a slight increase in branching, but a marked reduction in stem diameter, reduced internode elongation, and developmental delays in nodal system of adventitious root formation.
    22. Characterization knockout mutants of the cellulose-synthase-like-1 (csld1) gene showed a narrow-organ phenotype with 50% lower biomass accumulation arising mainly from ineffective cross-wall formation early in organogenesis and demonstrated that early-development partitioning of photosynthate to cell wall material can have significant impacts on biomass accumulation later in development.
    23. Insertional mutagenesis screens in Chlamydomonas cells grown under CO2-limiting conditions have identified new physical components of the carbon concentrating mechanism (CCM) including carbonic anhydrase enzymes and structural proteins essential for pyrenoid formation
    24. Understanding the mechanisms by which Chlamydomonas acclimates to low CO2 concentrations is important for evaluating the potential for increasing biomass production for biofuels from microalgal feedstocks as well as for transferring of all or part of this CCM into higher plants.
    25. Systematic mutagenesis studies of the carbonic anhydrase gene family in Arabidopsis will help identify the functional roles of this key enzyme in photosynthesis and improving crop yields in the future.
    26. Research into understanding the function of PTOX (plastid terminal oxidase), a versatile plastoquinol (terminal) oxidase in plastid membranes, might lead to novel strategies to manipulate and stabilize the photosynthetic capacity and quality of important crop plants during development and under stress conditions.
    27. Improvements in the properties of Rubisco are an obvious target for improving photosynthetic performance. However, without understanding its interactions with Rubisco activase, no such improvements are likely. Therefore, identifying the mechanism(s) for regulation of Rubisco activase and the roles of the two isoforms is necessary to ensure that modified Rubisco will function properly in the chloroplast.
    28. Research into understanding the mechanistic details of Rubisco activases exceptional instability at high temperature is essential for developing strategies to maintain plant yields under warmer temperatures.
    29. Engineered changes in either the large or small subunit of Rubisco far from the active site can influence carboxylation catalytic efficiency and CO2/O2 specificity. These regions might serve as targets for the design of an improved Rubisco using combinatorial approaches.
    30. Studies with hybrid Rubisco enzymes showed that the small subunit is responsible for targeting Rubisco to the algal pyrenoid. Thus, understanding the specific structural interactions required for establishing the pyrenoid will be critical to elucidating the details of CO2 concentrating mechanisms in algae.
    31. Characterization of essential nature of the AtDJ1C gene for Arabidopsis growth and development is the first example of any DJ-1 homolog being necessary for viability in any organism and provides a useful tool with which to investigate macromolecular interactions of this interesting superfamily of proteins.
    32. Production, in less than two years, of rice plants resistant to the major Asian plant disease, bacterial blight, using newly developed TAL Effector Nuclease (TALEN) technology for targeted knockout of the rice disease susceptibility gene, Os11N3.
    33. Publication of an extensive set of transcriptomics data showing that in Chlamydomonas CO2 deprivation causes massive changes in gene expression levels (i.e., >38% of genes are up regulated or down regulated by 2-fold or more) in less than three hours after the shift of cells from CO2 replete to CO2 deficient conditions.
    Last Modified: 29-Dec-2012
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